Anti-FANCA/FAA抗体(ab5063)

Reviews (3)Q&A (2)References (10)
Western blot - Anti-FANCA/FAA antibody (ab5063)

    Key features and details

    • Rabbit polyclonal to FANCA/FAA
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

    概述

    • 产品名称

      Anti-FANCA/FAA抗体
      参阅全部 FANCA/FAA 一抗

    • 描述

      兔多克隆抗体to FANCA/FAA

    • 宿主

      Rabbit

    • 特异性

      Detects a band at 130kDa in HeLa cell lysate that we believe corresponds to FANCA/FAA.

    • 经测试应用

      适用于: WBmore details

    • 种属反应性

      与反应: Human

    • 免疫原

      Synthetic peptide corresponding to Human FANCA/FAA aa 995-1009 conjugated to keyhole limpet haemocyanin.
      Sequence:

      SRSYDHSENSDLVFG-C


      Database link: O15360
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

    • 常规说明

       This product was previously labelled as FANCA

       

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    性能

    • 形式

      Liquid

    • 存放说明

      Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    • 存储溶液

      Preservative: 0.01% Sodium azide
      Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
    • Concentration information loading...

    • 纯度

      Immunogen affinity purified

    • 克隆

      多克隆

    • 同种型

      IgG

    • 研究领域

    应用

    The Abpromise guarantee

    Abpromise™承诺保证使用ab5063于以下的经测试应用

    “应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

    应用 Ab评论 说明
    WB (3)
    1/500 - 1/2000. Detects a band of approximately 130 kDa (predicted molecular weight: 163 kDa).
    说明
    WB
    1/500 - 1/2000. Detects a band of approximately 130 kDa (predicted molecular weight: 163 kDa).

    靶标

    • 功能

      DNA repair protein that may operate in a postreplication repair or a cell cycle checkpoint function. May be involved in interstrand DNA cross-link repair and in the maintenance of normal chromosome stability.

    • 疾病相关

      Defects in FANCA are a cause of Fanconi anemia (FA) [MIM:227650]. FA is a genetically heterogeneous, autosomal recessive disorder characterized by progressive pancytopenia, a diverse assortment of congenital malformations, and a predisposition to the development of malignancies. At the cellular level it is associated with hypersensitivity to DNA-damaging agents, chromosomal instability (increased chromosome breakage), and defective DNA repair.

    • 翻译后修饰

      Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation is required for the formation of the nuclear complex. Not phosphorylated in cells derived from groups A, B, C, E, F, G, and H.

    • 细胞定位

      Nucleus. Cytoplasm. The major form is nuclear. The minor form is cytoplasmic.
    • Information by UniProt

    • 数据库链接

      • 别名

        • FA 1 antibody
        • FA antibody
        • FA H antibody
        • FA1 antibody
        • FAA antibody
        • FACA antibody
        • FAH antibody
        • Fanca antibody
        • FANCA_HUMAN antibody
        • FANCH antibody
        • Fanconi anemia complementation group A antibody
        • Fanconi anemia complementation group H antibody
        • Fanconi anemia group A protein antibody
        • Fanconi anemia type 1 antibody
        • MGC75158 antibody
        • Protein FACA antibody
        see all

      图片

      • Western blot - Anti-FANCA/FAA antibody (ab5063)
        Western blot - Anti-FANCA/FAA antibody (ab5063)
        Anti-FANCA/FAA antibody (ab5063) at 1/1500 dilution + HeLa whole cell lysate at 35 µg

        Secondary
        IRDye 800 conjugated Goat anti-Rabbit IgG [H&L] at 1/10000 dilution

        Performed under reducing conditions.

        Predicted band size: 163 kDa
        Observed band size: 130 kDa why is the actual band size different from the predicted?
        Additional bands at: 42 kDa, 75 kDa. We are unsure as to the identity of these extra bands.

      文献 (10)

      发表研究结果有使用 ab5063？请让我们知道，以便我们可以引用本数据表中的参考文章。

      ab5063 被引用在 10 文献中.

      • Román-Rodríguez FJ  et al. NHEJ-Mediated Repair of CRISPR-Cas9-Induced DNA Breaks Efficiently Corrects Mutations in HSPCs from Patients with Fanconi Anemia. Cell Stem Cell 25:607-621.e7 (2019). PubMed: 31543367
      • Abdul-Sater Z  et al. FANCA safeguards interphase and mitosis during hematopoiesis in vivo. Exp Hematol 43:1031-1046.e12 (2015). IF . PubMed: 26366677
      • Vanuytsel K  et al. FANCA knockout in human embryonic stem cells causes a severe growth disadvantage. Stem Cell Res 13:240-50 (2014). WB . PubMed: 25108529
      • Rio P  et al. Targeted gene therapy and cell reprogramming in Fanconi anemia. EMBO Mol Med 6:835-48 (2014). WB ; Human . PubMed: 24859981
      • Shen C  et al. Regulation of FANCD2 by the mTOR pathway contributes to the resistance of cancer cells to DNA double-strand breaks. Cancer Res 73:3393-401 (2013). PubMed: 23633493
      • Meier D & Schindler D Fanconi anemia core complex gene promoters harbor conserved transcription regulatory elements. PLoS One 6:e22911 (2011). WB . PubMed: 21826217
      • Lu WT  et al. The Fanconi anemia pathway is downregulated upon macrophage differentiation through two distinct mechanisms. Cell Cycle 10: (2011). WB ; Human . PubMed: 21926477
      • Tulpule A  et al. Knockdown of Fanconi anemia genes in human embryonic stem cells reveals early developmental defects in the hematopoietic lineage. Blood 115:3453-62 (2010). WB ; Human . PubMed: 20089964
      • Mattera L  et al. The E1A-associated p400 protein modulates cell fate decisions by the regulation of ROS homeostasis. PLoS Genet 6:e1000983 (2010). WB ; Human . PubMed: 20548951
      • Wilson JB  et al. FANCG promotes formation of a newly identified protein complex containing BRCA2, FANCD2 and XRCC3. Oncogene 27:3641-52 (2008). PubMed: 18212739

      客户评价及客户问答

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      1-5 of 5 Abreviews or Q&A

      Question

      Anbei der vollständig ausgefüllte Fragebogen!



      Vielen dank!

      Mit besten grüßen

      1) Abcam product code ab 96089, ab 5063, ab54676, ab54645, ab 6264

      2) Abcam order reference number or product batch number

      3) Description of the problem: see pictures below, either to high cross reactivity or no signal detectable

      4) Sample preparation:
      Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysate
      Lysis buffer: RIPA
      Protease inhibitors: sigma (http://www.sigmaaldrich.com/catalog/product/sigma/p8340?lang=de®ion=AT)
      Phosphatase inhibitors: sigma
      Reducing agent
      Boiling for ≥5 min? yes/no yes
      Protein loaded ug/lane or cells/lane 15-100 µg
      Positive control yes
      Negative control

      5) Percentage of gel 10%
      Type of membrane Nylon
      Protein transfer verified yes (Ponseau red)
      Blocking agent and concentration 5% BSA in 1xTBS/T (as well as 5% milk in 1xTBS/T but in milk no signal could be detected)
      Blocking time 4 °C over night, as well as 1 h room temperature
      Blocking temperature see above

      6) Primary antibody (If more than one was used, describe in “additional notes”) :
      Concentration or dilution: FancG 1:500, FancA 1:1000, XPD 1:1000, CSB 1:1500
      Diluent buffer 5% BSA as well as milk in 1xTBS/T
      Incubation time over night
      Incubation temperature: 4°C

      7) Secondary antibody:
      Species:
      Reacts against:
      Concentration or dilution 1:1000
      Diluent buffer 5% BSA in 1x TBS/T
      Incubation time: 1h
      Incubation temperature: room temperature
      Fluorochrome or enzyme conjugate: HRP

      8) Washing after primary and secondary antibodies:
      Buffer 1x TBS/T
      Number of washes 3

      9)Detection method
      10) How many times have you run this staining?
      Do you obtain the same results every time?
      What steps have you altered to try and optimize the use of this antibody?
      * changed incubation time
      * changes milk to BSA
      * blocked over night or 1h

      Read More
      Answer

      Vielen Dank für Ihren Anruf und dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

      Es tut mir leid, dass Sie Probleme mit unseren Antikörper hatten.

      Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen, bzw noch ein paar zusätzliche Fragen stellen:

      1. Um was für Zelllinien handelt es sich hier, bzw aus welcher Spezies wurden die Zellen ursprünglich gewonnen? Sind die Zellen transfektiert?

      2. Was ist Ihre Positiv- Kontrolle?

      3.Verwenden Sie einen Anti-Mouse Sekundär- AKim falle der ab54645 und ab54676?

      4. Verwenden Sie immer einen frischen Blot pro Antikörper oder strippen Sie diesen zwischendurch?

      5. Haben Sie Ihre Zellen transfektiert oder wollen Sie das endogene Protein detektieren? Für den ersten Fall senken Sie den Proteingehalt auf 5- 10ug pro Bahn ab, im zweiten Fall gehen Sie runter auf 20 ug.

      6. Filtern Sie die Blockmittelllösung, dadurch sollten die schwarzen Punkte verschwinden und die Ergebnisse klarer werden.

      7. Senken Sie die Konzentration der Sekundär- Antikörper auf mindestens 1/5000 ab.

      8. Blockieren Sie weiterhin eine Stunde und inkubieren sie in der höchsten von uns empfohlenenVerdünnung über Nacht bei 4C.



      Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte und zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

      Read More

      Question

      long list of abs, two orders, only two work!
      works either not at all or strong cross-reactivity

      Read More
      Answer

      Vielen Dank für Ihren Anruf.

      Es tut mir leid zu hören, dass Sie Probleme mit diesen Antikörpern haben.

      Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.


      Ich freue mich, bald wieder von Ihnen zu hören.

      Read More

      Western blot abreview for Anti-FANCA/FAA antibody

       Below Average
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (HeLa)
      Gel Running Conditions
      Reduced Denaturing
      Loading amount
      50 µg
      Specification
      HeLa
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      提交于 Aug 14 2007

      Western blot abreview for Anti-FANCA/FAA antibody

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (THP1 - Human acute monocytic leukemia cell)
      Loading amount
      150 µg
      Specification
      THP1 - Human acute monocytic leukemia cell
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
      Read More

      Dr. Wei-Ting Lu

      Verified customer

      提交于 Jul 24 2007

      Western blot abreview for Anti-FANCA/FAA antibody

       Average
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (Cal51 & SW620)
      Loading amount
      750 cells
      Specification
      Cal51 & SW620
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
      Read More

      Mr. Richard Allum

      Verified customer

      提交于 Oct 24 2006

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